Composition for accelerating change in muscle type

ABSTRACT

The present invention provides a composition for accelerating a change in muscle type, increasing the amount of muscle, strengthening muscle, enhancing athletic abilities, reducing lipids, suppressing the accumulation of lipids, lowering blood sugar, controlling body weight or lowering body weight, containing a PPAR-δ promoter, an AMPK promoter and a PGC1-α promoter as an active ingredient

CROSS REFERENCE TO RELATED APPLICATION

This application is a divisional application of U.S. application Ser.No. 13/638,813, filed Oct. 1, 2012, and is incorporated in its entiretyby reference herewith.

TECHNICAL FIELD

The present disclosure relates to a composition for accelerating achange in muscle type.

BACKGROUND ART

Muscles generated through aerobic exercise consume body fats by usingfats rather than carbohydrates as primary energy source. Since lesscarbohydrate is consumed, the blood sugar level is maintained and lesshunger is experienced. In addition, since ATP is produced from fatsrather than from carbohydrates, less reactive oxygen species areproduced during the ATP generation and cell damage by the reactiveoxygen species can be prevented. However, not all exercise exhibits sucheffect. Only aerobic exercises such as running and bicycling can providesuch effect since they change the fast-twitch type muscles intoslow-twitch type.

Human muscle consists of fast-twitch (type II) muscles and slow-twitch(type I) muscles approximately half-and-half. The fast-twitch musclelooks white since it lacks myoglobin and uses carbohydrates as primaryenergy source. This type of muscle is mainly formed through anaerobicexercise such as weight training. It has large muscle mass and is usedfor quick motion. In contrast, the slow-twitch muscle looks red becauseof myoglobin and uses fats instead of carbohydrates as primary energysource. It is mainly formed through aerobic exercise, has small musclemass and is used for slow, long motion. The slow-twitch muscles enableexercise for a long time since they are rich in mitochondria and havehigh mitochondrial activity and thus resistance to fatigue is improved.

In general, sprinting athletes have fast-twitch muscles and slow-twitchmuscles at a proportion of about 70:30 and have well-developed upperbodies. In contrast, marathoners usually have fast-twitch muscles andslow-twitch muscles at a proportion of about 30:70 and tend to have slimbodies. Marathoners are undersized than non-athletes of the same bodyweight (fast-twitch muscle:slow-twitch muscle=about 50:50) becausemuscles are about 5 times smaller in volume than fats of the sameweight. To have a body of a marathoner through aerobic exercise, oneshould take exercise of 65-75% intensity based on the maximum heart ratefor over 6 weeks, at least 3 times a week, more than 40 minutes a day,which is not so easy.

DISCLOSURE Technical Problem

The present disclosure is directed to providing a composition foraccelerating a change in muscle type, increasing the amount of muscle,strengthening muscle, enhancing athletic ability, reducing lipids,suppressing the accumulation of lipids, lowering blood sugar,controlling body weight or lowering body weight.

Technical Solution

In one general aspect, the present disclosure provides a composition foraccelerating a change in muscle type, increasing the amount of muscle,strengthening muscle, enhancing athletic ability, reducing lipids,suppressing the accumulation of lipids, lowering blood sugar,controlling body weight or lowering body weight, comprising a peroxisomeproliferator-activated receptor-δ (PPAR-δ) activating substance, anAMP-activated protein kinase (AMPK) activating substance and aperoxisome proliferator-activated receptor gamma coactivator 1-α(PGC1-α) activating substance as an active ingredient.

Advantageous Effects

The composition according to an aspect of the present disclosure, whichcomprises a peroxisome proliferator-activated receptor-δ (PPAR-δ)activating substance, an AMP-activated protein kinase (AMPK) activatingsubstance and a peroxisome proliferator-activated receptor gammacoactivator 1-α (PGC1-α) activating substance as an active ingredient,exhibits an effect of accelerating a change in muscle type, increasingthe amount of muscle, strengthening muscle, enhancing athletic ability,reducing lipids, suppressing the accumulation of lipids, lowering bloodsugar, controlling body weight or lowering body weight.

DESCRIPTION OF DRAWINGS

FIG. 1 shows fluorescence intensity of 22 natural product extracts as ameasure of binding to LBD of PPAR-δ.

FIG. 2 shows the level of AMPK phosphorylation when muscle cells weretreated for 24 hours with Salvia miltiorrhiza, Angelica gigas,eriobotryae folium, Gynostemma pentaphyllum leaf, Citrus unshiu orHouttuynia cordata extract.

FIG. 3 shows the level of luciferase expression as a result ofactivation of the PGC1-α activating substance by several natural productextracts including astragali radix extract.

FIG. 4 shows a result of treating muscle cells with various substancesincluding 200 μg/mL Artemisia princeps extract, EPA, Gynostemmapentaphyllum leaf extract, astragali radix extract or mixtures thereofand comparing expression level of the CPT1β, PDK4 and PGC1-α genes witha negative control group (GAA: Gynostemma pentaphyllum leafextract+Artemisia princeps extract+astragali radix extract, GEA:Gynostemma pentaphyllum leaf extract+EPA+astragali radix extract, GEP:Gynostemma pentaphyllum leaf extract+EPA+pinotin (pine nut extract),CAA: Citrus unshiu extract+Artemisia princeps extract+astragali radixextract, CEA: Citrus unshiu extract+EPA+astragali radix extract, CEP:Citrus unshiu extract+EPA+pinotin (pine nut extract), GAP: Gynostemmapentaphyllum leaf extract+Artemisia princeps extract+pinotin (pine nutextract), CAP: Citrus unshiu extract+Artemisia princeps extract+pinotin(pine nut extract), ChEP: chitooligosaccharide+EPA+pinotin (pine nutextract), REP: resveratrol+EPA+pinotin (pine nut extract), ChAA:chitooligosaccharide+Artemisia princeps extract+astragali radix extract,ChEA: chitooligosaccharide+EPA+astragali radix extract).

FIG. 5 shows exercise duration of groups to which Artemisia princepsextract, EPA, Gynostemma pentaphyllum leaf extract, astragali radixextract or mixtures thereof were administered.

FIG. 6 shows body weight increase of groups to which Artemisia princepsextract, EPA, Gynostemma pentaphyllum leaf extract, astragali radixextract or mixtures thereof were administered.

FIG. 7 shows epididymal fat weight of groups to which Artemisia princepsextract, EPA, Gynostemma pentaphyllum leaf extract, astragali radixextract or mixtures thereof were administered.

FIG. 8 shows plantaris muscle weight of groups to which Artemisiaprinceps extract, EPA, Gynostemma pentaphyllum leaf extract, astragaliradix extract or mixtures thereof were administered.

FIG. 9 compares the amount of adiponectin and insulin in groups to whichArtemisia princeps extract, EPA, Gynostemma pentaphyllum leaf extract,astragali radix extract or mixtures thereof were administered relativeto a negative control.

FIG. 10 shows muscle type of groups to which Artemisia princeps extract,EPA, Gynostemma pentaphyllum leaf extract, astragali radix extract ormixtures thereof were administered.

FIG. 11 shows hepatic triglyceride level of groups to which Artemisiaprinceps extract, EPA, Gynostemma pentaphyllum leaf extract, astragaliradix extract or mixtures thereof were administered.

FIG. 12 shows the size of lipid droplets in brown adipose tissue ofgroups to which Artemisia princeps extract, EPA, Gynostemma pentaphyllumleaf extract, astragali radix extract or mixtures thereof wereadministered.

FIG. 13 compares the expression level of the CPT1β, PDK4 and PGC1-αgenes in the muscle cells of groups to which Artemisia princeps extract,EPA, Gynostemma pentaphyllum leaf extract, astragali radix extract ormixtures thereof were administered relative to a negative control.

MODE FOR INVENTION

As used herein, “extract” means a substance extracted from a naturalproduct, regardless of extraction method or ingredients. The term isused in a broad sense including, for example, ingredients soluble inwater or an organic solvent extracted from a natural substance using thesolvent, or specific ingredients of a natural substance such as oilextracted therefrom.

As used herein, “metabolism” means the processes of breakdown ofnutrients taken up by an organism, production of substances or energyrequired for life activities, and transport of unnecessary substancesout of the body. When the metabolism occurs actively, energy consumptionin the body increases.

Hereinafter, the present disclosure is described in further detail.

Peroxisome proliferator-activated receptor-δ (PPAR-δ) is a factorexpressed in muscle, brown adipose tissue, etc. It is reported thatadiposity is inhibited in mouse in which this transcription factor isoverexpressed as a result of accelerated oxidation of fatty acids in fatcells. Also, it is known that an activator of PPAR-δ promotes metabolismof skeletal muscle cells, improves sensitivity to insulin, reduces fatcells and inhibits inflammatory response by increasing the expression ofsuch proteins as carnitine palmitoyltransferase Iβ (CPT1β), pyruvatedehydrogenase kinase isozyme 4 (PDK4), etc.

AMP-activated protein kinase (AMPK) is a protein activated when AMP issensed in cells. The protein protects cells from external stress byinhibiting the ATP-consuming signaling pathway and activating theATP-synthetizing signaling pathway. It is known to inhibit fat synthesisand promote fatty acid oxidation in muscles and to suppress sugarproduction in the liver by inhibiting gluconeogenesis.

However, only with a substance that activates PPAR-δ and AMPK, theeffect of changing muscle type from the fast-twitch type to theslow-twitch type or increasing the slow-twitch type muscle is notsignificant. In particular, the effect is only slight in adults whosemuscles are already developed. Thus, only with a substance thatactivates PPAR-δ and AMPK, it is difficult to achieve the aerobicexercise effect of increasing the amount of muscle, strengtheningmuscle, reducing body fat, suppressing the accumulation of lipids,lowering blood sugar, controlling body weight or lowering body weight,or the like.

During aerobic exercise, the amount of mitochondria which produce energyincreases in muscles. Mitochondria oxidize fatty acids and ATP isproduced as the fatty acid oxidation is accelerated by mitochondria. Thenumber and capacity of mitochondria that oxidize fatty acids areregulated by peroxisome proliferator-activated receptor gammacoactivator 1-α (PGC1-α).

In an aspect, the present disclosure provides a composition foraccelerating a change in muscle type, increasing the amount of muscle,strengthening muscle, enhancing athletic ability, reducing lipids,suppressing the accumulation of lipids, lowering blood sugar,controlling body weight or lowering body weight, comprising a PPAR-δactivating substance, an AMPK activating substance and a PGC1-αactivating substance as an active ingredient.

The PPAR-δ activating substance increases expression of enzymespromoting fat and sugar metabolism in muscle cells by binding to PPAR-δand increasing its activity. The AMPK activating substance may regulateenergy metabolism by accelerating phosphorylation of the AMPK protein.The PGC1-α activating substance may increase mitochondrial biosynthesisby acting on the PGC1-α promoter and promoting expression of the PGC1-αgene.

In an exemplary embodiment of the present disclosure, a compositioncomprising the PPAR-δ activating substance, the AMPK activatingsubstance and the PGC1-α activating substance as an active ingredientmay accelerate change in muscle type. The change in muscle type includesa change from a fast-twitch type to a slow-twitch type or an increase inthe slow-twitch type. As described above, one having a large amount ofslow-twitch type muscles looks slimmer even when the body weight is thesame since the slow-twitch muscle has a smaller volume than thefast-twitch muscle. Further, the slow-twitch muscle reduces lipids andsuppresses accumulation of the lipids in the body since it uses fatsinstead of carbohydrates as primary energy source. In addition, sincethe slow-twitch muscle is used for slow, long motion as compared to thefast-twitch muscle, the increase in slow-twitch muscle may improveendurance.

In an exemplary embodiment of the present disclosure, a compositioncomprising the PPAR-δ activating substance, the AMPK activatingsubstance and the PGC1-α activating substance as an active ingredientmay improve athletic ability and metabolism by increasing the amount ofmuscle in the body and strengthening muscle. Also, it provides theeffect of reducing lipids in the body and blood and lowering blood sugarby increasing expression of the genes involved in fat and sugarmetabolism in muscle cells such as CPT1β, PDK4, PGC1α, GAPDH, etc.

Accordingly, when the composition comprising the PPAR-δ activatingsubstance, the AMPK activating substance and the PGC1-α activatingsubstance as an active ingredient is used for an animal or human, it mayprovide an effect similar to that of aerobic exercise and may allowcontrol and lowering of body weight.

In an exemplary embodiment of the present disclosure, the PPAR-δactivating substance included in the composition comprises one or moreof Artemisia princeps extract, eicosapentaenoic acid (EPA), black pepper(piperis nigri fuctus) extract, green mate (Ilex paraguariensis) extractand puerariae radix (root of a plant in the genus Pueraria) extract. TheEPA is a naturally-occurring PPAR-δ ligand.

In an exemplary embodiment of the present disclosure, the AMPKactivating substance included in the composition comprises one or moreof Gynostemma pentaphyllum leaf extract, Citrus unshiu extract andHouttuynia cordata extract.

In an exemplary embodiment of the present disclosure, the PGC1-αincluded in the composition comprises one or more of astragali radixextract, puerariae flos (flower of a plant in the genus Pueraria)extract and Citrus leiocarpa extract.

In an exemplary embodiment of the present disclosure, the extracts maybe obtained by extracting the corresponding natural products accordingto a commonly employed method. In another exemplary embodiment of thepresent disclosure, the extracts may be obtained by heating andextracting the corresponding natural products in an organic solventincluding water or alcohol, and then filtering and concentrating underreduced pressure. In another exemplary embodiment of the presentdisclosure, the organic solvent may be a C₁-C₅ low alcohol, although notbeing limited thereto. For example, the C₁-C₅ low alcohol may be one ormore selected from a group consisting of methanol, ethanol, isopropylalcohol, n-propyl alcohol, n-butanol and isobutanol.

In an exemplary embodiment of the present disclosure, the compositioncomprising the PPAR-δ activating substance, the AMPK activatingsubstance and the PGC1-α activating substance as an active ingredientmay comprise the activating substance, the AMPK activating substance andthe PGC1-α activating substance in an amount of 1-80 wt % based on thetotal weight of the composition. In another exemplary embodiment of thepresent disclosure, the PPAR-δ p activating substance, the AMPKactivating substance and the PGC1-α activating substance may be includedin an amount of 5-60 wt % based on the total weight of the composition.In another exemplary embodiment of the present disclosure, the PPAR-δactivating substance, the AMPK activating substance and the PGC1-αactivating substance may be included in an amount of 10-30 wt % based onthe total weight of the composition. When the PPAR-δ activatingsubstance, the AMPK activating substance and the PGC1-α activatingsubstance are included in an amount of 1-80 wt % based on the totalweight of the composition, the effect desired by the present disclosurecan be achieved while ensuring both stability and safety of thecomposition and satisfying cost effectiveness.

In an exemplary embodiment of the present disclosure, in the compositioncomprising the PPAR-δ activating substance, the AMPK activatingsubstance and the PGC1-α activating substance as an active ingredient,each of the PPAR-δ activating substance, the AMPK activating substanceand the PGC1-α activating substance may be included in an amount of 1-30wt % based on the total weight of the composition. In another exemplaryembodiment of the present disclosure, each of the PPAR-δ activatingsubstance, the AMPK activating substance and the PGC1-α activatingsubstance may be included in an amount of 5-20 wt % based on the totalweight of the composition. In another exemplary embodiment of thepresent disclosure, each of the PPAR-δ activating substance, the AMPKactivating substance and the PGC1-α activating substance may be includedin an amount of 5-10 wt % based on the total weight of the composition.When each of the PPAR-δ activating substance, the AMPK activatingsubstance and the PGC1-α activating substance is included in an amountof 1-30 wt % based on the total weight of the composition, the effectdesired by the present disclosure can be achieved while ensuring bothstability and safety of the composition and satisfying costeffectiveness.

In an exemplary embodiment of the present disclosure, in the compositioncomprising the PPAR-δ activating substance, the AMPK activatingsubstance and the PGC1-α activating substance as an active ingredient,the PPAR-δ activating substance, the AMPK activating substance and thePGC1-α activating substance may be mixed at a ratio of 1-10:1-10:1-10.In another exemplary embodiment of the present disclosure, the PPAR-δactivating substance, the AMPK activating substance and the PGC1-αactivating substance may be mixed at a ratio of 1-5:1-5:1-5. In anotherexemplary embodiment of the present disclosure, the PPAR-δ activatingsubstance, the AMPK activating substance and the PGC1-α activatingsubstance may be mixed at a ratio of 1-2:1-2:1-2. When the PPAR-δactivating substance, the AMPK activating substance and the PGC1-αactivating substance are mixed at the above-described ratio, the effectdesired by the present disclosure can be achieved while ensuring bothstability and safety of the composition and satisfying costeffectiveness.

The composition according to the present disclosure is applicable notonly to animals but also to human.

In an aspect, the present disclosure provides a pharmaceuticalcomposition comprising the PPAR-δ activating substance, the AMPKactivating substance and the PGC1-α activating substance as an activeingredient.

The pharmaceutical composition may further comprise a pharmaceuticaladjuvant such as antiseptic, stabilizer, hydrating agent, emulsifyingaccelerator, salt and/or buffer for controlling osmotic pressure, etc.or other therapeutically useful substance, and may be prepared intovarious formulations for oral or parenteral administration.

The formulation for oral administration may include, for example,tablet, pill, hard or soft capsule, liquid, suspension, emulsion, syrup,powder, dust, granule, pellet, or the like. These formulations maycomprise, in addition to the active ingredient, a surfactant, a diluent(e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose orglycine) or a lubricant (e.g., silica, talc, stearic acid and magnesiumor calcium salt thereof or polyethylene glycol). The tablet may comprisea binder such as magnesium aluminum silicate, starch paste, gelatin,tragacanth, methyl cellulose, sodium carboxymethyl cellulose andpolyvinylpyrrolidone, and may occasionally comprise a pharmaceuticaladditive such as a disintegrant, e.g. starch, agar, alginic acid or asodium salt thereof, an absorbent, a colorant, a flavor, a sweetener, orthe like. The tablet may be prepared according to the commonly employedmixing, granulation or coating method.

The formulation for parenteral administration may include, for example,injection, drop, ointment, lotion, gel, cream, spray, suspension,emulsion, suppository, patch, etc., but is not limited thereto.

In an exemplary embodiment of the present disclosure, the pharmaceuticalcomposition may be administered orally or parenterally, e.g. rectally,topically, transdermally, intravenously, intramuscularly,intraabdominally, subcutaneously, etc.

A pharmaceutically acceptable amount, i.e. administration dose, of theactive ingredient will vary depending on the age, sex and body weight ofthe subject to be treated, particular disease or pathological conditionto be treated, severity of the disease or pathological condition,administration route and discretion of the prescriber. Determination ofthe administration dose considering these factors is within the level ofthose skilled in the art. A general administration dose may be 0.01-2000mg/kg/day, specifically 1-100 mg/kg/day. However, the describedadministration dose does not limit the scope of the present disclosureby any means.

In another aspect, the present disclosure provides a food compositioncomprising the PPAR-δ activating substance, the AMPK activatingsubstance and the PGC1-α activating substance as an active ingredient.The food composition may be a health food composition.

The formulation of the food or health food composition is notparticularly limited. For example, it may be prepared into tablet,granule, drink, caramel, diet bar, tea bag, etc. Each formulation of thefood composition may comprise, in addition to the active ingredient,other ingredients commonly used in the art. Those other ingredients maybe selected by those skilled in the art without difficulty byconsidering the particular formulation or purpose of use and may providesynergic effect.

Determination of the dose of the active ingredient is within the levelof those skilled in the art. A daily dose may vary depending on variousfactors including the age, physical condition, complication, etc. of thesubject.

In another aspect, the present disclosure provides a beauty carecomposition comprising the PPAR-δ activating substance, the AMPKactivating substance and the PGC1-α activating substance as an activeingredient.

For example, the beauty care composition may be a cosmetic composition.The cosmetic composition may comprise a cosmetologically ordermatologically allowable medium or base. It may be provided in anytopically applicable form including, for example, solution, gel, solid,anhydrous slurry, oil-in-water emulsion, water-in-oil emulsion,multiemulsion, suspension, microemulsion, microcapsule, microgranule,ionic (liposome) or non-ionic vesicular dispersion, foam, or an aerosolcomposition comprising a pressurized propellant. Such compositions maybe prepared by a method commonly employed in the art.

The beauty care composition may further comprise fatty substance,organic solvent, solubilizer, thickening agent, gelling agent, softeningagent, antioxidant, suspending agent, stabilizer, foaming agent,aromatic, surfactant, water, ionic or non-ionic emulsifier, filler,sequestrant, chelating agent, preservative, vitamin, blocker, hydratingagent, essential oil, dye, pigment, hydrophilic or lipophilic activeagent, lipid vesicle or other adjuvant commonly used in the field ofcosmetics or dermatology. The adjuvant is added in an amount commonlyused in the field of cosmetics or dermatology.

The beauty care composition is not particularly limited with regard tothe formulation thereof and the formulation may be determinedappropriately depending on purposes. For example, the beauty carecomposition may be provided as one or more formulation selected from agroup consisting of toilet water, lotion, essence, cream, ointment, gel,pack, patch, spray, powder foundation, emulsion foundation, concealstick, hand or foot lotion, hand or foot cream, hand or foot oil, handor foot essence, hand or foot cleanser, soap, cleansing cream, cleansinglotion, cleansing foam and cleansing water, but is not limited thereto.

The features and effects of the present disclosure will be described indetail through preparation examples, examples and test examples.However, the following preparation examples, examples and test examplesare provided for illustrative purposes only and are not intended tolimit the scope of the present disclosure.

Preparation Example 1 Preparation of Artemisia princeps Extract

Artemisia princeps cultivated in Chengchen-dong, Jecheon-si,Chungcheongbuk-do, Korea was purchased. After adding 70% ethanol (3 L)to Artemisia princeps (300 g), the mixture was stirred at 70-80° C. for3 hours. After repeating this procedure 2 times, a filtrate obtained byfiltering through filter paper was concentrated under reduced pressureusing a rotary vacuum evaporator, which was freeze-dried to obtain drypowder (29 g).

Preparation Example 2 Preparation of Gynostemma pentaphyllum LeafExtract

Gynostemma pentaphyllum leaf (1 kg) was crushed and repeatedly extracted2-3 times for 12-24 hours at 90-100° C. after adding about 10 volumeequivalents of water or alcohol. After filtering, the filtrate wasconcentrated under reduced pressure to obtain Gynostemma pentaphyllumleaf extract.

Preparation Example 3 Preparation of Astragali Radix Extract

Astragali radix cultivated in Korea was purchased from a local market.Astragali radix and triply distilled water were mixed at a ratio of 1:10and extraction was performed for 3 hours using an extractor. Afterrepeating this procedure 2 times, a filtrate obtained by filteringthrough filter paper was concentrated under reduced pressure using arotary vacuum evaporator, which was freeze-dried to obtain astragaliradix extract as dry powder.

EXAMPLES

The Artemisia princeps extract, the Gynostemma pentaphyllum leaf extractand the astragali radix extract prepared above were mixed at a ratio of1:1:1 (Example 1). Also, EPA, the Gynostemma pentaphyllum leaf extractand the astragali radix extract were mixed at a ratio of 1:1:1 (Example2).

Test Example 1 Screening of PPAR-δ Activating Substance

Peroxisome proliferator-activated receptor-δ (PPAR-δ) is activated whena ligand binds to the ligand binding domain (LBD) of the protein andregulates expression of other genes. Accordingly, a substance shouldeasily bind to the LBD to activate the transcription factor. Based onthis fact, about 90 edible natural products were screened for PPAR-δactivating substance.

The natural products were dissolved in dimethyl sulfoxide (DMSO) to 200,100 or 50 μg/mL and the degree of binding to the LBD of PPAR-δ wasquantified with fluorescence intensity using the LanthaScreen TR-FRETperoxisome proliferator receptor delta coactivator assay kit(Invitrogen, CA). DMSO was used as negative control. The result for the22 natural products that showed superior binding to the LBD of PPAR-δ isshown in FIG. 1.

As seen from FIG. 1, Artemisia princeps, black pepper, green mate andpuerariae radix showed superior activity among them. In particular,Artemisia princeps had an EC₅₀ value for the LBD of PPAR-δ ligand of 9μg/mL.

Test Example 2 Screening of AMPK Activating Substance

About 90 edible natural products were screened for AMP-activated proteinkinase (AMPK) activating effect as follows.

Immature C2C12 cells were purchased from the American Tissue CultureCollection (ATCC; USA). The cells were cultured in Dulbecco's modifiedEagle's Medium (DMEM; Gibco 1210-0038) containing 10% fetal bovine serum(FBS) in a 5% CO₂ incubator until 70% confluency while replacing themedium every other day. The cells were induced to differentiate intomuscle cells in a medium containing 2% horse serum (HS). After culturingfor 4 days in a medium containing 2% HS, the muscle cells were treatedwith the natural products dissolved in DMSO (200 μg/mL) for 24 hours.

As positive control, 1 mL of AICAR (Cell Signaling Technology, Inc., UK)which is well known as AMPK activator was used. As negative control,DMSO of a volume of 1/1000 as that of the medium was used. 24 hourslater, the cells were washed with phosphate buffered saline (PBS),treated with 200 μL of a protein extraction buffer containing 8 M urea,2% 3-[(3-chloramidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS),50 mM dithiothreitol (DTT), 2 M thiourea, 2 mM phenylmethanesulfonylfluoride (PMSF) and 100 μg/μL leupeptin, and then kept at roomtemperature for 10 minutes. Subsequently, after centrifugation at 4° C.and 15,000 g for 10 minutes and collecting the supernatant, protein wasquantitated using Bio-Rad Protein Dye Reagent™. 100 g of protein wasseparated based on size with 8% SDS-PAGE and blotted onto PDF membrane(Bio-Rad) at 50 V for 12 hours. The blots obtained were blocked with 5%skim milk for 1 hour, reacted with anti-AMPK-α (Cell signaling),anti-phospho-AMPK-α (Cell Signaling) and anti-6-actin as primaryantibody and with a secondary antibody conjugated with horse radishperoxidase (Amersham Biosciences), and detected using the enhancedchemiluminescence (ECL) kit (Amersham Biosciences). The reacted blotswere exposed to Fuji X-ray film and developed to identify the degree ofprotein expression. Bands on the film were scanned with PowerLook 2100XL (UMAX) and analyzed using the ImageMaster 2D Elite program (AmershamBioscience).

The result for 6 plants that exhibited superior AMPK activating effectamong the tested natural products is shown in FIG. 2. From FIG. 2, itcan be seen that the cells treated with Gynostemma pentaphyllum leaf,Citrus unshiu and Houttuynia cordata show remarkably increased AMPKphosphorylation.

Test Example 3 Screening of PGC1-α Activating Substance

APRDC PGC1-α promoter cells were used to test PGC1-α promoter activatingeffect of 90 natural products. The APRDC PGC1-α promoter cells are humanhepatic cells (KCTC 11218BP) in which a vector with the PGC1-α promoterand the luciferase gene genetically fused is stably expressed.

First, APRDC PGC1-α promoter cells were treated with the 90 naturalproducts dissolved in DMSO (200 μg/mL) for 24 hours. Then, after washing2 times with PBS, the activity of the reporter gene luciferase wasmeasured using the Steady Glo luciferase assay kit (Promega, Cat No.E2520). The luciferase activity was measured by transferring the sampleto a 96-well plate and detecting fluorescence using a luminometer. DMSOwas used as negative control.

The result for several natural products that exhibited superior APRDCPGC1-α promoter activating effect among the tested natural products isshown in FIG. 3. As seen from FIG. 3, treatment with astragali radix,puerariae flos or Citrus leiocarpa resulted in stronger fluorescence ascompared to the negative control DMSO or other substances. This meansthat astragali radix, puerariae flos and Citrus leiocarpa have superiorPGC1-α promoter activating effect.

Test Example 4 Evaluation of Gene Expression in Muscle Cells (atCellular Level)

The effect of treatment with a 1:1:1 mixture of the Artemisia princepsextract, the Gynostemma pentaphyllum leaf extract and the astragaliradix extract, which were confirmed to activate PPAR-δ, AMPK and PGC1-α,on the expression of the genes involved in fat and sugar metabolism inmuscle cells was investigated.

Immature C2C12 muscles were purchased from the ATCC (USA). The cellswere cultured in DMEM (Gibco 1210-0038) containing 10% FBS in a 5% CO₂incubator until 70% confluency while replacing the medium every otherday. The cells were induced to differentiate into muscle cells in amedium containing 2% HS. After culturing for 4 days in a mediumcontaining 2% HS, the muscle cells were treated with the Artemisiaprinceps extract, the Gynostemma pentaphyllum leaf extract, theastragali radix extract or mixtures thereof (200 μg/mL). As negativecontrol, DMSO of a volume of 1/1000 as that of the medium was used.After culturing the cells treated with each sample for 24 hours at 37°C. and washing 2 times with cold saline, RNA was extracted using theTRIzol reagent (Invitrogen). cDNA was synthesized using the extractedand quantitated RNA (1 μg/μL) and a reverse transcription system(Promega).

Expression profile of the CPT1β, PDK4, PGC1α and GAPDH genes wasmeasured using the synthesized cDNA and primers and probes (AppliedBiosystems; CPT1β, Mm00487200_m1; PDK4, Mm00447181_m1; PGC1-αMm00447181_m1; GAPDH, Mm99999915_q1) designed for the genes. PCR andanalysis were performed using the Rotor-Gene 3000 system (CorbettResearch, Sydney, Australia). The result is shown in FIG. 4.

As seen from FIG. 4, treatment with the mixture of the three substancesincreased the expression of the CPT1β, PDK4 and PGC1-α genes 2 times ormore as compared to when the negative control substance was used or thesubstance was used alone. Accordingly, it can be seen thatadministration of a mixture of the substances that activate the PPAR-δ,AMPK and PGC1-α respectively can promote fat and sugar metabolism inmuscle.

Test Example 5 Evaluation of Exercise Duration

(1) Preparation of Experimental Mouse

5-week-old male C57BL/6 mice were divided into a normal diet group, ahigh-fat diet group and an aerobic exercise group, with 10 mice pergroup. The normal diet group was fed with D12450B (10% fat, ResearchDiets, Inc., NJ, USA) and the high-fat diet group was fed with D12492(60% fat) for 8 weeks. The aerobic exercise group was given the high-fatdiet and made to run on a treadmill for 40 minutes at 15 m/min, 5 timesa week. The test groups were given the high-fat diet and orallyadministered with Gynostemma pentaphyllum leaf extract, Artemisiaprinceps extract, astragali radix extract, EPA, a mixture of Artemisiaprinceps extract, Gynostemma pentaphyllum leaf extract and astragaliradix extract or a mixture of EPA, Gynostemma pentaphyllum leaf extractand astragali radix extract (200 mg/kg) once a day. Body weight and feedintake were measured every week.

(2) Measurement of Exercise Duration

8 weeks after the administration of the test substances, theexperimental mice were forced to run on a treadmill at 15 m/min andexercise duration was measured. The result is shown in FIG. 5. As seenfrom FIG. 5, the group to which the mixture of Gynostemma pentaphyllumleaf extract, Artemisia princeps extract and astragali radix extract orthe mixture of Gynostemma pentaphyllum leaf extract, EPA and astragaliradix extract was administered showed about 70% increased exerciseduration as compared to the untreated negative control group or thegroups to which the test substances were administered alone.

Accordingly, it can be seen that administration of a mixture of thesubstances that activate PPAR-δ, AMPK and PGC1-α respectively enablesexercise for a long time.

Test Example 6 Measurement of Body Weight and Tissue Weight

5-week-old male C57BL/6 mice were grouped with 10 mice per group. Theywere orally administered with Gynostemma pentaphyllum leaf extract,Artemisia princeps extract, astragali radix extract or EPA alone, amixture of Artemisia princeps extract, Gynostemma pentaphyllum leafextract and astragali radix extract or a mixture of EPA, Gynostemmapentaphyllum leaf extract and astragali radix extract (200 mg/kg) astest substances once a day. The experimental mice were fasted for 12hours before autopsy.

FIG. 6 shows change in body weight for 8 weeks. As seen from FIG. 6, thegroup to which the mixture of Gynostemma pentaphyllum leaf extract,Artemisia princeps extract and astragali radix extract and the group towhich the mixture of Gynostemma pentaphyllum leaf extract, EPA andastragali radix extract was administered showed 26% and 28% decreasedbody weight, respectively, as compared to the untreated negative controlgroup or the groups to which the test substances were administeredalone.

Epididymal fat weight is shown in FIG. 7. As seen from FIG. 7, theweight decreased in the group to which the mixture of Gynostemmapentaphyllum leaf extract, Artemisia princeps extract and astragaliradix extract and the group to which the mixture of Gynostemmapentaphyllum leaf extract, EPA and astragali radix extract wasadministered by 34% and 36%, respectively, as compared to the untreatednegative control group or the groups to which the test substances wereadministered alone.

The result of separating and weighing the mouse plantaris muscle isshown in FIG. 8. As seen from FIG. 8, the plantaris muscle weightincreased in the group to which the mixture of Gynostemma pentaphyllumleaf extract, Artemisia princeps extract and astragali radix extract andthe group to which the mixture of Gynostemma pentaphyllum leaf extract,EPA and astragali radix extract was administered by about 55% ascompared to the untreated negative control group or the groups to whichthe test substances were administered alone.

Accordingly, it can be seen that administration of a mixture of thesubstances that activate PPAR-δ, AMPK and PGC1-α respectively increasesthe amount of muscle while lowering body weight and epididymal fatweight.

Test Example 7 Blood Analysis

5-week-old male C57BL/6 mice were grouped with 10 mice per group. Theywere orally administered with Gynostemma pentaphyllum leaf extract,Artemisia princeps extract, astragali radix extract or EPA alone, amixture of Artemisia princeps extract, Gynostemma pentaphyllum leafextract and astragali radix extract or a mixture of EPA, Gynostemmapentaphyllum leaf extract and astragali radix extract (200 mg/kg) astest substances for 8 weeks, once a day. Then, blood analysis wasperformed to evaluate blood sugar level and serum triglyceride andcholesterol levels.

The blood sugar level was measured using the Accu-Check Active kit(Roche Diagnostics, Seoul, Korea).

The blood sample was centrifuged at 3000 rpm for 10 minutes to separateserum. Triglyceride and cholesterol levels in the separated serum wereanalyzed using the Vitalab Selectra E analyzer (Vital Scientific,Dieren, The Netherlands). The result is shown in Table 1.

TABLE 1 Blood sugar Triglyceride Cholesterol Group (mg/dL) (mg/dL)(mg/dL) Control 183.5 ± 9.1  100.4 ± 8.7 160.2 ± 5.33 Gynostemma 180.2 ±8.47 102.98 ± 9.7  167.4 ± 9.7  pentaphyllum leaf extract Artemisiapriceps extract 176.3 ± 10.1 99.34 ± 5.4 170.3 ± 11.2 Astragali radixextract 179.9 ± 8.92 89.74 ± 5.4 159.3 ± 4.5  EPA 181.2 ± 10.9 101.23 ±11.3 164.9 ± 12.1 Gynostemma 169.3 ± 8.6   62.5 ± 8.5 175.34 ± 10.23pentaphyllum leaf extract + Artemisia priceps extract + astragali radixextract Gynostemma 169.6 ± 6.5   80.8 ± 8.3 156.2 ± 5.4  pentaphyllumleaf extract + EPA + astragali radix extract

As seen from Table 1, the group to which the mixture of Gynostemmapentaphyllum leaf extract, Artemisia princeps extract and astragaliradix extract or the mixture of Gynostemma pentaphyllum leaf extract,EPA and astragali radix extract was administered showed significantlylower blood sugar and triglyceride levels as compared to the untreatednegative control group or the groups to which the test substances wereadministered alone.

Accordingly, it can be seen that administration of a mixture of thesubstances that activate PPAR-δ, AMPK and PGC1-α respectively providesthe effect of lowering blood sugar and triglyceride levels.

Test Example 8 Analysis of Adiponectin and Insulin Levels

5-week-old male C57BL/6 mice were grouped with 10 mice per group. Theywere orally administered with Gynostemma pentaphyllum leaf extract,Artemisia princeps extract, astragali radix extract or EPA alone, amixture of Artemisia princeps extract, Gynostemma pentaphyllum leafextract and astragali radix extract or a mixture of EPA, Gynostemmapentaphyllum leaf extract and astragali radix extract (200 mg/kg) astest substances for 8 weeks, once a day. Then, adiponectin and insulinlevels were measured.

Blood was taken in an anticoagulant tube and centrifuged at 3000 rpm for10 minutes to separate serum. The serum was diluted 20,000 times and thequantity of expressed adiponectin was measured using the MouseAdiponectin Quantikine kit (R&D Systems). Insulin level was measuredusing the Mouse Insulin Quantikine kit (R&D Systems) after diluting theserum 20,000 times. The result is shown in FIG. 9.

The group to which the mixture of Gynostemma pentaphyllum leaf extract,Artemisia princeps extract and astragali radix extract or the mixture ofGynostemma pentaphyllum leaf extract, EPA and astragali radix extractwas administered showed about 50% increased adiponectin level ascompared to the untreated negative control group or the groups to whichthe test substances were administered alone. Also, the group to whichthe mixture of Gynostemma pentaphyllum leaf extract, Artemisia princepsextract and astragali radix extract and the group to the mixture ofGynostemma pentaphyllum leaf extract, EPA and astragali radix extractwas administered showed 60% and 55% decreased insulin level,respectively, as compared to the untreated negative control group or thegroups to which the test substances were administered alone.

Accordingly, it can be seen that administration of a mixture of thesubstances that activate PPAR-δ, AMPK and PGC1-α respectively mayincrease the level of the protein hormone adiponectin, which is involvedin the regulation of blood sugar and fatty acid metabolism, and decreaseinsulin level. Decreased insulin level means that blood sugar ismaintained low.

Test Example 9 Analysis of Change in Muscle Type

5-week-old male C57BL/6 mice were grouped with 10 mice per group toevaluate change in muscle type after administration of test substances.The experimental mice were orally administered with Gynostemmapentaphyllum leaf extract, Artemisia princeps extract, astragali radixextract or EPA alone, a mixture of Artemisia princeps extract,Gynostemma pentaphyllum leaf extract and astragali radix extract or amixture of EPA, Gynostemma pentaphyllum leaf extract and astragali radixextract (200 mg/kg) as test substances for 8 weeks, once a day. The micewere fasted for 12 hours before autopsy.

Plantaris muscle was taken from the mice of each test group and frozenin liquid nitrogen. The frozen tissue was sliced to 10-μm thicknessusing a cryotome, fixed in acetone for 10 minutes, and washed 3 timeswith PBS. After blocking for 2 hours with PBS containing 10% FBS, thetissue sections were treated with anti-slow-twitch myosin antibody(Abcam, Cambridge, UK) as primary antibody and with fluoresceinisothiocyanate (FITC)-conjugated anti-mouse IgG antibody (MolecularProbes) as secondary antibody. After mounting with antifade solution,the sections were examined by confocal microscopy.

The analysis result is shown in FIG. 10 and Table 2.

TABLE 2 Slow-twitch Fast-twitch Group muscle (%) muscle (%) Control 25.4± 1.6 74.5 ± 2.6 Gynostemma pentaphyllum leaf extract 30.6 ± 2.1 69.4 ±3.1 Artemisia priceps extract 24.3 ± 2.7  75.7 ± 3.76 Astragali radixextract 24.5 ± 1.9 75.5 ± 3.1 EPA 29.4 ± 4.2 70.6 ± 3.6 Gynostemmapentaphyllum leaf extract + 49.5 ± 3.4 44.3 ± 4.3 Artemisia pricepsextract + astragali radix extract Gynostemma pentaphyllum leaf extract +50.4 ± 4.3 55.6 ± 4.3 EPA + astragali radix extract

The group to which the mixture of Gynostemma pentaphyllum leaf extract,Artemisia princeps extract and astragali radix extract or the mixture ofGynostemma pentaphyllum leaf extract, EPA and astragali radix extractwas administered showed about 2 times increased slow-twitch myosin inthe muscle tissue as compared to the untreated negative control group orthe groups to which the test substances were administered alone. Thismeans that administration of the mixture of Gynostemma pentaphyllum leafextract, Artemisia princeps extract and astragali radix extract or themixture of Gynostemma pentaphyllum leaf extract, EPA and astragali radixextract changes mouse muscle type to the slow-twitch muscle. Also, thismeans that administration of a mixture of the substances that activatePPAR-δ, AMPK and PGC1-α respectively may provide a similar effect asthat of aerobic exercise in terms of change in muscle type.

Test Example 10 Analysis of Triglyceride in Hepatic Tissue

5-week-old male C57BL/6 mice were grouped with 10 mice per group. Theexperimental mice were orally administered with Gynostemma pentaphyllumleaf extract, Artemisia princeps extract, astragali radix extract or EPAalone, a mixture of Artemisia princeps extract, Gynostemma pentaphyllumleaf extract and astragali radix extract or a mixture of EPA, Gynostemmapentaphyllum leaf extract and astragali radix extract (200 mg/kg) astest substances for 8 weeks, once a day, and then triglyceride level inhepatic tissue was evaluated. The mice were fasted for 12 hours beforeautopsy.

Hepatic tissue was taken from the experimental mice and frozen in liquidnitrogen. The frozen tissue was sliced to 10-μm thickness using acryotome, reacted with 0.5% Oil Red 0 (0-0625, Sigma-Aldrich, USA) inpropylene glycol (031301, Samchun pure chemical Co. Ltd., Korea) for 8minutes at 60° C., and then reacted for 2 minutes with 85% propyleneglycol. Thereafter, the tissue sections were washed for 1 minute withflowing water, counter-stained with Mayer's hematoxylin (10029273, DAKO,USA), and observed under an optical microscope. The result is shown inFIG. 11.

As seen from FIG. 11, the group to which the mixture of Gynostemmapentaphyllum leaf extract, Artemisia princeps extract and astragaliradix extract or the mixture of Gynostemma pentaphyllum leaf extract,EPA and astragali radix extract was administered looks lighter ascompared to the untreated negative control group or the groups to whichthe test substances were administered alone, suggesting thattriglycerides are remarkably decreased in hepatic tissue.

Accordingly, it can be seen that administration of a mixture of thesubstances that activate PPAR-δ, AMPK and PGC1-α respectively may reducetriglycerides in hepatic tissue.

Test Example 11 Analysis of Lipid Droplets in Brown Adipose Tissue

5-week-old male C57BL/6 mice were grouped with 10 mice per group. Theexperimental mice were orally administered with Gynostemma pentaphyllumleaf extract, Artemisia princeps extract, astragali radix extract or EPAalone, a mixture of Artemisia princeps extract, Gynostemma pentaphyllumleaf extract and astragali radix extract or a mixture of EPA, Gynostemmapentaphyllum leaf extract and astragali radix extract (200 mg/kg) astest substances for 8 weeks, once a day, and then lipid droplets inbrown adipose tissue was examined. The mice were fasted for 12 hoursbefore autopsy.

Brown adipose tissue was taken from each test group and fixed in 10%neutral formalin. Following paraffin embedding, the paraffin-embeddedtissue was sliced to 3-μm thickness using a microtome, stained withhematoxylin (10029273, DAKO, USA) and eosin (075K4346, Sigma-Aldrich,USA) (H&M), and observed under an optical microscope. The result isshown in FIG. 12.

As seen from FIG. 12, the group to which the mixture of Gynostemmapentaphyllum leaf extract, Artemisia princeps extract and astragaliradix extract or the mixture of Gynostemma pentaphyllum leaf extract,EPA and astragali radix extract was administered showed lipid dropletsof remarkably decreased size in the brown adipose tissue as compared tothe untreated negative control group or the groups to which the testsubstances were administered alone.

Accordingly, it can be seen that administration of a mixture of thesubstances that activate PPAR-δ, AMPK and PGC1-α respectively may reducethe size of lipid droplets in brown adipose tissue.

Test Example 12 Evaluation of Gene Expression in Muscle Cells(Individual Level)

5-week-old male C57BL/6 mice were grouped with 10 mice per group. Theexperimental mice were orally administered with Gynostemma pentaphyllumleaf extract, Artemisia princeps extract, astragali radix extract or EPAalone, a mixture of Artemisia princeps extract, Gynostemma pentaphyllumleaf extract and astragali radix extract or a mixture of EPA, Gynostemmapentaphyllum leaf extract and astragali radix extract (200 mg/kg) astest substances for 8 weeks, once a day, and then gene expression levelin muscle cells was evaluated. The mice were fasted for 12 hours beforeautopsy.

Muscle taken from each test group was dissected and RNA was extractedusing the TRIzol reagent (Invitrogen). cDNA was synthesized using theextracted and quantitated RNA (1 μg/μL) and a reverse transcriptionsystem (Promega). Expression profile of the CPT1β, PDK4, PGC1α and GAPDHgenes was measured using the synthesized cDNA and primers and probes(Applied Biosystems; CPT1β, Mm00487200_m1; PDK4, Mm00447181_m1; PGC1-αMm00447181_m1; GAPDH, Mm99999915_q1) designed for the genes. PCR andanalysis were performed using the Rotor-Gene 3000 system (CorbettResearch, Sydney, Australia). The result is shown in FIG. 13.

As seen from FIG. 13, the group to which the mixture of Gynostemmapentaphyllum leaf extract, Artemisia princeps extract and astragaliradix extract or the mixture of Gynostemma pentaphyllum leaf extract,EPA and astragali radix extract was administered showed about 2 timesincreased expression of the genes as compared to the untreated negativecontrol group or the groups to which the test substances wereadministered alone. Accordingly, it can be seen that administration of amixture of the substances that activate PPAR-δ, AMPK and PGC1-αrespectively may promote fat and sugar metabolism in muscle.

Formulation examples of the pharmaceutical composition, food compositionand beauty care composition according to the present disclosurecomprising the PPAR-δ activating substance, the AMPK activatingsubstance and the PGC1-α activating substance as an active ingredientare described hereinafter. However, the scope of the present disclosureis not limited to the following examples.

Formulation Example 1 Preparation of Tablet

Example 1 or Example 2 50 mg Corn starch 100 mg Lactose 100 mg Magnesiumstearate 2 mg Vitamin C 50 mg

The above ingredients are mixed and prepared into a tablet according toa commonly employed method.

Formulation Example 2 Preparation of Capsule

Example 1 or Example 2 50 mg Corn starch 100 mg Lactose 100 mg Magnesiumstearate 2 mg Vitamin C 50 mg Serine 50 mg

The above ingredients are mixed and filled in a gelatin capsule toprepare a capsule according to a commonly employed method.

Formulation Example 3 Preparation of Liquid

Example 1 or Example 2 100 mg High-fructose corn syrup 10 g Mannitol 5 gVitamin C 50 mg Serine 50 mg Fat adequate Purified water balance

According to a commonly employed method, the above ingredients aredissolved by adding to purified water. After adding an adequate amountof lemon flavor, the ingredients are mixed and purified water is addedto make a total volume 100 mL. The mixture is filled in a brown bottleand sterilized to prepare a liquid.

Formulation Example 4 Preparation of Health Food

Example 1 or Example 2 1000 mg Vitamin mixture Vitamin A acetate 70 μgVitamin E 1.0 mg Vitamin B₁ 0.13 mg Vitamin B₂ 0.15 mg Vitamin B₆ 0.5 mgVitamin B₁₂ 0.2 μg Vitamin C 10 mg Biotin 10 μg Nicotinamide 1.7 mgFolic acid 50 μg Calcium pantothenate 0.5 mg Mineral mixture Ferroussulfate 1.75 mg Zinc oxide 0.82 mg Magnesium carbonate 25.3 mg Potassiumphosphate monobasic 15 mg Calcium phosphate dibasic 55 mg Potassiumcitrate 90 mg Calcium carbonate 100 mg Magnesium chloride 24.8 mg

The compositions of the vitamin and mineral mixtures are presented as anexample appropriate for health food. However, the compositions can bevaried differently. According to a commonly employed method, the aboveingredients are mixed, prepared into granule and then used forpreparation of a health food composition.

Formulation Example 5 Preparation of Drink

Example 1 or Example 2 1000 mg Citric acid 1000 mg Oligosaccharide 100 gPlum concentrate 2 g Taurine 1 g Purified water to make 1000 mL

According to a commonly employed method, the above ingredients are mixedand heated at 85° C. for about 1 hour while stirring. Thus preparedsolution is filtered, collected in a sterilized 2-L container, sealed,and then stored in a refrigerator, for use in preparation of a drinkcomposition.

Formulation Example 6 Preparation of Cream

A cream is prepared using the ingredients described in Table 3 accordingto a commonly employed method.

TABLE 3 Ingredients wt % Example 1 or Example 2 2.0 Polysorbate 60 1.5Sorbitan sesquiolate 0.5 PEG60 hydrogenated castor oil 2.0 Liquidparaffin 10 Squalane 5.0 Caprylic/capric triglyceride 5.0 Glycerin 5.0Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Antisepticadequate Pigment adequate Flavor adequate Purified water to 100

1-2. (canceled)
 3. A method for increasing the amount of muscle,strengthening muscle or enhancing athletic ability, comprisingadministering an effective amount of a peroxisome proliferator-activatedreceptor-δ (PPAR-δ) activating substance, an AMP-activated proteinkinase (AMPK) activating substance and a peroxisomeproliferator-activated receptor gamma coactivator 1-α (PGC 1-α)activating substance to a subject in such need to increase the amount ofmuscle, strengthen muscle or enhance athletic ability.
 4. A method forreducing lipids, suppressing the accumulation of lipids or loweringblood sugar, comprising administering an effective amount of aperoxisome proliferator-activated receptor-δ (PPAR-δ) activatingsubstance, an AMP-activated protein kinase (AMPK) activating substanceand a peroxisome proliferator-activated receptor gamma coactivator 1-α(PGC1-α) activating substance to a subject in such need to reducelipids, suppress the accumulation of lipids or lower blood sugar.
 5. Amethod for controlling body weight or lowering body weight, comprisingadministering an effective amount of a peroxisome proliferator-activatedreceptor-δ (PPAR-δ) activating substance, an AMP-activated proteinkinase (AMPK) activating substance and a peroxisomeproliferator-activated receptor gamma coactivator 1-α (PGC 1-α)activating substance to a subject in such need to control body weight orlower body weight. 6-9. (canceled)
 10. The method according to claim 3,wherein the PPAR-δ activating substance comprises one or more ofArtemisia princeps extract, eicosapentaenoic acid (EPA), black pepperextract, green mate extract and puerariae radix extract.
 11. The methodaccording to claim 4, wherein the PPAR-δ activating substance comprisesone or more of Artemisia princeps extract, eicosapentaenoic acid (EPA),black pepper extract, green mate extract and puerariae radix extract.12. The method according to claim 5, wherein the PPAR-δ activatingsubstance comprises one or more of Artemisia princeps extract,eicosapentaenoic acid (EPA), black pepper extract, green mate extractand puerariae radix extract.
 13. The method according to claim 3,wherein the AMPK activating substance comprises one or more ofGynostemma pentaphyllum leaf extract, Citrus unshiu extract andHouttuynia cordata extract.
 14. The method according to claim 4, whereinthe AMPK activating substance comprises one or more of Gynostemmapentaphyllum leaf extract, Citrus unshiu extract and Houttuynia cordataextract.
 15. The method according to claim 5, wherein the AMPKactivating substance comprises one or more of Gynostemma pentaphyllumleaf extract, Citrus unshiu extract and Houttuynia cordata extract. 16.The method according to claim 3, wherein the PGC 1-α activatingsubstance comprises one or more of astragali radix extract, puerariaeflos extract and Citrus leiocarpa extract.
 17. The method according toclaim 4, wherein the PGC 1-α activating substance comprises one or moreof astragali radix extract, puerariae flos extract and Citrus leiocarpaextract.
 18. The method according to claim 5, wherein the PGC 1-αactivating substance comprises one or more of astragali radix extract,puerariae flos extract and Citrus leiocarpa extract.
 19. The methodaccording to claim 3, wherein the PPAR-δ activating substance, the AMPKactivating substance and the PGC1-α activating substance are mixed at aratio of 1-10:1-10:1-10.
 20. The method according to claim 4, whereinthe PPAR-δ activating substance, the AMPK activating substance and thePGC1-α activating substance are mixed at a ratio of 1-10:1-10:1-10. 21.The method according to claim 5, wherein the PPAR-δ activatingsubstance, the AMPK activating substance and the PGC1-α activatingsubstance are mixed at a ratio of 1-10:1-10:1-10.